Dna intercalator detection

ABSTRACT

A DNA intercalator detection system can include a filtration unit a control sample conditioner operably coupled with the filtration unit and an analytic unit operably coupled with the filtration unit and control sample conditioner and having an electronic chemical array (ECA) reaction component. A data processing unit is operably coupled with the analytic unit and configured to compare and determine a difference between electronic data of a test sample and a conditioned control sample from the ECA reaction component. The difference provides an indication of whether or not a DNA intercalator is present in the test sample.

BACKGROUND

DNA intercalators contain many chemical substances that have a harmfuleffect on human health. Typical examples of these substances includecarcinogens and endocrine disruptors, which have a polycyclic aromaticmolecular structure. For instance, testing systems have been developedto monitor benzopyrene and dioxins, which are typical DNA intercalators.In one system, samples collected in the environment (e.g., atmosphere,river, and soil) are taken to a laboratory and subjected to a robustchemical analysis. In another system, gene expression profiles for cellssensitive to DNA intercalators are detected in a laboratory setting.There is an interest for the creation of new risk-screening systems inwhich the level of a DNA intercalator in the environment can be checkedon site easily and quickly in advance of implementing laboratorytesting.

SUMMARY

In one aspect, a DNA intercalator detection system can include afiltration unit; a control sample conditioner unit configured to receivea portion of a filtered sample from the filtration unit and remove a DNAintercalator from the sample so as to produce a conditioned controlsample; an analytic unit configured to receive a portion of the filteredsample from the filtration unit (e.g., test sample) and to receive theconditioned control sample from the control sample conditioner unit,wherein the analytic unit has one or more electronic chemical arrays(ECAs); and a data processing unit configured to compare and determine adifference between electronic current of the filtered sample and theconditioned control sample from the one or more ECAs. The differenceprovides an indication of whether or not a DNA intercalator is presentin the filtered sample.

In one aspect, a method for detecting a DNA intercalator in anenvironmental sample can include filtering an environmental sample toobtain a filtered sample; splitting the filtered sample into two or moreseparate portions; conditioning one or more portions of the filteredsample such that one or more DNA intercalators are removed therefrom toform a conditioned control sample; contacting a portion of the filteredsample with double stranded nucleic acids having a strand linked to anECA, wherein the portion of the filtered sample contacted with thedouble stranded nucleic acids is a test sample; contacting theconditioned control sample with double stranded nucleic acids having astrand linked to an ECA; obtaining electronic data from an electroniccurrent of the test sample and from an electronic current of theconditioned control sample; and determining a difference between theelectronic data from the test sample and the conditioned control sample.The difference provides an indication of whether or not a DNAintercalator is present in the sample.

In one aspect, a method for detecting a DNA intercalator in anenvironmental sample can include introducing the environmental sampleinto a portable DNA intercalator detection system at the location,wherein the DNA intercalator detection system can include a filtrationunit, a control sample conditioner unit, an analytic unit, and a dataprocessing unit; filtering the environmental sample with the filtrationunit so as to remove environmental substances from the environmentalsample and provide a filtered sample; conditioning a portion of thefiltered sample with the control sample conditioner unit so as to removeone or more DNA intercalators and form a conditioned control sample;contacting, in the analytic unit, the test sample and conditionedcontrol sample with double stranded nucleic acids having a strand linkedto an ECA; obtaining electronic data from an electronic current of thetest sample and the conditioned control sample when in contact with thedouble stranded nucleic acids; and determining a difference between theelectronic data from the test sample and the conditioned control samplewith the data processing unit. The difference provides an indication ofwhether or not a DNA intercalator is present in the sample.

The foregoing summary is illustrative only and is not intended to be inany way limiting. In addition to the illustrative aspects, embodiments,and features described above, further aspects, embodiments, and featureswill become apparent by reference to the drawings and the followingdetailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is an illustrative example of a DNA intercalator testing system.

FIG. 2 is an illustrative example of a DNA intercalator testing system.

FIG. 3 is an illustrative example of an analytic unit.

FIG. 4 is an illustrative example of a method of performing DNAintercalator testing with the system described herein.

FIG. 5 is an illustrative example of an electrochemical array (ECA)analytic module.

FIGS. 6A-6B are illustrative examples of an electrochemical array (ECA)analytic module.

FIGS. 7A-7B are illustrative examples of an electrochemical array (ECA)analytic module.

FIGS. 8A-8B are illustrative examples of a portable DNA intercalatortesting system.

DETAILED DESCRIPTION

In the following detailed description, reference is made to theaccompanying drawings, which form a part hereof. In the drawings,similar symbols typically identify similar components, unless contextdictates otherwise. The illustrative embodiments described in thedetailed description, drawings, and claims are not meant to be limiting.Other embodiments may be utilized, and other changes may be made,without departing from the spirit or scope of the subject matterpresented herein. It will be readily understood that the aspects of thepresent disclosure, as generally described herein, and illustrated inthe Figures, can be arranged, substituted, combined, separated, anddesigned in a wide variety of different configurations, all of which areexplicitly contemplated herein.

A testing system can be used in testing various environmentalcompositions (e.g., water) for the presence or levels of DNAintercalators, such as benzo[a]pyrene and dibenzo-p-dioxin as well asother intercalators. DNA intercalators intercalate or reversibly bindwith double stranded DNA. Intercalation occurs when molecules of anappropriate size and chemical nature fit between base pairs of DNA. DNAintercalators are known to be a health risk and identification of thepresence and/or determining the amount or concentration of DNAintercalators in the environment can be important. The testing systemcan have various components that can process a sample for testing, whichcomponents are described in more detail below. The testing system can beinstalled in a laboratory or configured to be portable.

The DNA intercalators can be hydrophobic DNA intercalators, such as butnot limited to DNA intercalators having one or more aromatic rings,polyaromatics, heterocyclic aromatics, and combinations thereof, such aspolycyclic aromatic hydrocarbons. The flat portions of these types ofsubstances intercalate between base pairs of DNA. Non-limiting examplesof hydrophobic DNA intercalators includes carbazoles, acridines,anthracenes, anthracyclines, ethidium bromides, proflavines,daunomycins, doxorubicins, thalidomides, quinacrines, derivativesthereof, and combinations thereof.

In one option, the testing system is portable. As such, the testingsystem can be delivered to a testing site or environmental location.Examples of portability can include the testing system having a sizethat one or more persons can move the testing system, and may include adeck having the components of the system. The portable system can alsoinclude wheels for moving and transportation of the system. In light ofthe rising public concern over environmental and health issues, thetesting system can quickly be taken to a test site to test for thepresence or amounts of DNA intercalators. As such, the portable testingsystem can be contained within a vehicle, on trailer, or the like fortransportation. The portable testing system can then perform an onsiteanalysis of the environment for risk-screening for DNA intercalators.This portable testing system has advantages over conventional equipmentand methods which require samples to be transported to a specificlaboratory for a robust chemical analysis by a professional technician.

In one embodiment, the testing system can be configured to perform arough study or general analysis for the presence or amount of one ormore potential DNA intercalators. After the rough study, a positiveresult can then qualify the sample as being suitable a robust chemicalanalysis for an in-depth analysis of the DNA intercalator. On the otherhand, the testing system can be configured to identify the presence oramount accurately enough to be a standalone system and analyticalprocess.

The DNA intercalator detection system can include one or more of afiltration unit, a control sample conditioner unit, an analytic unit,and a data processing unit. The control sample conditioner unit can beconfigured to receive a portion of a filtered sample from the filtrationunit. The analytic unit can be configured to receive a portion of thefiltered sample from the filtration unit and a conditioned controlsample from the control sample conditioner unit. Also, the analytic unitcan have one or more electronic chemical arrays (ECAs). For example, theanalytic unit can have one ECA for the filtered sample and one ECA forthe conditioned control sample. Alternatively, a single ECA can be usedto analyze the filtered sample and the conditioned control sample. Thedata processing unit can be configured to receive electronic data fromthe analytic unit, and can be configured to compare and determine adifference between electronic data of the filtered sample (e.g., a testsample) and the conditioned control sample. The difference in electronicdata between the filtered sample and the conditioned control sample canprovide an indication of whether or not a DNA intercalator is present inthe test sample.

The filtration unit can be configured to filter components from asample. The filtration unit can include a filter that can be configuredas any type of filter that can allow certain components of a sample topass through the filter while trapping or retaining certain componentsfrom the sample. The filtration unit can include a filter that canseparate environmental and/or biological materials from an environmentalsample. In an embodiment for filtering water samples, the filter can bea membrane filter having a hydrophilic property. Alternatively, thefilter can include a hydrophobic property. Examples of filters caninclude without limitation mixed cellulose esters (MCE), glass filtermaterials, nylon, cellulose acetate, polyvinylidene difluoride (PVDF),polytetrafluoroethylene (PTFE), or polycarbonate.

In one embodiment, the testing system can include a sample dividerconfigured to receive a filtered sample from the filtration unit andthen to split the filtered sample into two or more separatecompositions. One of the compositions can be a test sample and one ormore of the separate compositions being the control sample that isprocessed into a conditioned control sample as described herein.

The control sample conditioner unit can include a hydrophobic componentconfigured to attract and retain hydrophobic DNA intercalators from thecontrol sample. The control sample conditioner unit can be configured asa separation column (e.g., FPLC, HPLC) or other separation unit that hasa hydrophobic component that can attract hydrophobic DNA intercalatorsfrom the sample by hydrophobic interaction. For example, the hydrophobiccomponent can include without limitation C8 to C20 hydrocarbons that arelinear, cyclic, or branched, substituted or unsubstituted, aromatic ornon-aromatic, carbon homo atoms or hetero atoms, such as phenylcomponents and octadecylsilyl-based components such as but not limitedto octadecylsilyl-silica gels, octadecylsilyl columns, andoctadecylsilyl matrices, and/or hydrocarbon components such as but notlimited to polymethacrylates having butyl, ether, or phenyl ligand.

The analytic unit is configured to receive samples and analyze thesamples for the presence of a hydrophobic DNA intercalator. Accordingly,the analytic unit includes sample receiving components and sampleanalyzing components, which are described in more detail below. Theanalytic unit can include any one or more components or configurations,some of which include: a port fluidly coupled with the sample dividerand configured to receive the test sample; a port fluidly coupled withthe control sample conditioner unit and configured to receive theconditioned control sample; one or more sample chambers configured toreceive one or more samples; one or more fluid pathways configured todeliver the test sample and conditioned control sample to the one ormore sample chambers; an integrated or removable ECA reaction componenthaving: a working electrode, counter electrode, and/or referenceelectrode (e.g., two electrode or three electrode detection system); anucleic acid strand operably coupled with one of the electrodes suchthat the electrodes can receive electrons from the nucleic acid; and acomplementary nucleic acid strand either hybridized or hybridizable withthe electrode-coupled nucleic acid; electronic components configured tomeasure an electronic current passed from the electrode-coupled nucleicacid; voltammetry electronic components; electronic componentsconfigured to obtain electronic data from the electrodes; a transmitterto transmit electronic data to the data processing unit; or a receiverto receive instructions from the data processing unit.

The data processing unit can include hardware and/or software configuredto operate components of the system, such as the filtration unit, sampledivider unit, control sample conditioner unit, or analytical unit. Thedata processing unit can also be configured to compare electronic datafrom the test sample with the conditioned control sample. As such, thedata processing unit can include a computer-readable memory devicehaving computer-executable instructions to operate the system and/oranalyze data obtained from testing samples with the system. The dataprocessing unit can be configured or programmed to automatically receivedata from the analytic unit and/or automatically process the data todetermine the presence of a DNA intercalator. Alternatively, the dataprocessing unit can be configured or programmed to receive instructionsfrom a user for the receipt or processing of data from the analyticunit.

The system can include one or more user interfaces, which include a userinput interface and an output interface. The user input interface caninclude common computing devices that allow a user to interface with acomputing system. Examples include keyboards, mice, light pens, touchscreens, buttons, knobs, switches, or the like. The output interface caninclude common computing devices that provide information to a user.Examples include monitors, screens, speakers, lights, printers, and thelike. Also, the system can be operably coupled with a communicationnetwork (e.g., wired, optical, or wireless) so that the data and/orresults of the processed data can be transmitted over the communicationnetwork.

A method for detecting a DNA intercalator in an environmental sample caninclude filtering an environmental sample so as to remove environmentalsubstances from the environmental sample and to provide a filteredsample; splitting the filtered sample into two or more separatecompositions, one or more of the separate compositions being a testsample and one or more of the separate compositions being a controlsample; conditioning the control sample so as to remove one or more DNAintercalators and to form a conditioned control sample; contacting thetest sample and conditioned control sample with double stranded nucleicacids each having a strand linked to an ECA; detecting electronic dataof the test sample and the conditioned control sample; determining adifference between the electronic data from the test sample and theconditioned control sample. The difference provides an indication ofwhether or not a DNA intercalator is present in the sample.

The method for detecting a DNA intercalator in an environmental samplecan also include any one or more of the following: collecting theenvironmental sample from a location and performing the method at thelocation; removing, with a filtration unit, biological materials fromthe environmental sample; contacting the control sample to a hydrophobiccomponent capable of attracting and retaining hydrophobic DNAintercalators from the control sample; measuring the electronic currentsof the test sample and conditioned control sample; hybridizing a nucleicacid strand coupled to the ECA with its complementary nucleic acid;conducting a voltammetry protocol; transmitting the electronic data to adata processing unit. For example, this can include processing theelectronic data in the data processing unit in order to determine adifference between the electronic data from the test sample and theconditioned control sample.

According to another aspect, a method can include determining an amountor health risk of a DNA intercalator or detecting a DNA intercalator inan environmental sample. Such a method can include: collecting anenvironmental sample from a location; introducing the environmentalsample into a DNA intercalator detection system (e.g., portable system)at the location; filtering the environmental sample with the filtrationunit so as to remove environmental substances from the environmentalsample and provide a filtered sample; conditioning a portion of thefiltered sample with the control sample conditioner unit so as to removeone or more DNA intercalators and form a conditioned control sample;contacting, separately in the analytic unit, the test sample andconditioned control sample with double stranded nucleic acids eachhaving a strand linked to a electronic chemical array (ECA); obtainingelectronic data from the electronic currents of the test sample and theconditioned control sample when contacted with the nucleic acids; anddetermining a difference between the electronic data from the testsample and the conditioned control sample with the data processing unit.The difference provides an indication of whether or not a DNAintercalator is present in the sample.

FIG. 1 depicts an illustrative example of a configuration of the DNAintercalator testing system 10, which includes a filtration unit 12, acontrol sample conditioner unit 14, an analytic unit 16, and a dataprocessing unit 18.

The filtration unit 12 can be configured to filter a sample, which isthen provided to the control sample conditioner unit 14. For example,fluid pathways, pipettes, or the like can provide the filtered sample tothe control sample conditioner unit 14. Similarly, the filtration unit12 and control sample conditioner unit 14 can provide samples to theanalytic unit 16. The analytic unit 16 can be in communication with thedata processing unit 18 so that data can be passed therebetween. Forexample, the analytic unit 16 can be in communication with the dataprocessing unit 18 by wires, optics, or wireless communication.

FIG. 2 depicts another illustrative example of a configuration of theDNA intercalator detection system 20, which includes a filtration unit12, a sample divider 22, a control sample conditioner unit 14, ananalytic unit 16, and a data processing unit 18.

In one embodiment, the testing system 10 or 20 does not include thefiltration unit 12 because samples can be obtained that do not need tobe filtered. For example, some water samples may not need filtration.

Similar to the embodiment of the DNA intercalator detection system 10 ofFIG. 1, the DNA intercalator detection system 20 of FIG. 2 includes thecomponents being operable with each other. As such, the filtration unit12 can provide a filtered sample to the sample divider 22 by using fluidpathways, pipettes, or the like.

The filtration unit 12 can be configured to receive an environmentalsample and to remove environmental substances from the environmentalsample and to provide a filtered sample. As such, the filtration unit 12can receive various types of samples ranging from soil, plantcomponents, animal components, water and the like. The filtration unit12 can include filter components, such as a filter, tubing, filterhousing, or the like (not shown), which are common in filtration units.Filtration units are common devices used to separate materials, and assuch any appropriate type of filtration unit can be included in the DNAintercalator detection system 10. The filtration unit 12 can beconfigured to separate environmental and/or biological materials from anenvironmental sample by capturing and retaining environmental and/orbiological materials from the sample.

In one embodiment, the filtration unit 12 may be configured to removematerials from the environmental sample, such as foreign matter,plankton, microbes, proteins, and the like. The filtration unit 12 caninclude a filter (not shown), which is configured to allow certainsubstances in a sample to pass therethrough while trapping and retainingsome substances from the sample. The filter can be formed from variousmaterials that are commonly used for filters: such as polymers such asbut not limited to polyvinylidene difluoride (PVDF), polyethylene,polypropylene, polytetrafluoroethylene (PTFE), or polycarbonate; metalssuch as but not limited to stainless steel, aluminum, or galvanizedsteel; ceramics such as but not limited to aluminum oxides, zirconia,carbides, borides, nitrides, or silicides; or composites such as but notlimited to woven wire with fiber metal felt and composite materialsformed from polymers, metals, and/or ceramics. For example, thefiltration unit 12 can include a filter made from polyvinylidenedifluoride (PVDF). Thus, the filtration unit 12 can be configured toremove foreign materials from the environmental sample while leaving theDNA intercalators in the filtered sample so as to improve theeffectiveness of the testing system and testing protocols.

The filtration unit 12 can include a removable filter that can bereplaced or replenished when the filtering capacity is reduced. In someinstances the substances, such as environmental or biological substancescan build up on the filter and reduce the ability of the filtration unit12 to function as a filter, which causes a reduced filtering capacity.The filter can be replaced as needed to retain an optimum or desirablefiltering capacity.

When included (e.g., in FIG. 2), a sample divider 22 can be any deviceor system configured to split a sample into two portions or to removesome portion of a sample from the bulk sample. The sample divider 22 caninclude a wide range of components to facilitate splitting a sample intotwo or more portions, such as pipettes, valves, fluid conduits andnetworks, reservoirs, chambers, cutting devices, spatulas, tongs, pumps,and the like. Such a sample divider 22 can be configured to split asolid, paste, gel, liquid or other sample format. The sample divider 22can be configured to split the filtered sample into two or more separatecompositions, one or more of the separate compositions being a testsample and one or more of the separate compositions being a controlsample. The sample divider 22 can be operably coupled with thefiltration unit 12 to receive a filtered sample, and then operablycoupled to the control sample conditioner unit 14 and the analytic unit16. For example, the sample divider 22 can be operably coupled with thefiltration unit 12 by fluid pathways that can move a sample from thefiltration unit 12 to the sample divider 22. A pipetting system can alsooperably couple the filtration unit 12 to the sample divider 22 by beingconfigured to pipette a sample from the filtration unit 12 to the sampledivider 22. The sample divider 22 can be similarly associated with thecontrol sample conditioner unit 14.

The sample divider 22 can be configured to split the filtered sampleinto two compositions at various ratios. The sample splitter can beconfigured to split the sample into a ratio of 9:1, 4:1, 7:3, 3:2, or1:1 with respect to the portion sent to the analytic unit 16 compared tothe control sample conditioner unit 14. However, other ratios can beused.

The control sample conditioner unit 14 can be configured to remove oneor more DNA intercalators from the control sample so as to form acleansed control sample. This allows a comparison between the filteredenvironmental sample with and without the DNA intercalators so that thepresence or amount of the DNA intercalators can be detected in theanalytic unit 16. The control sample conditioner unit 14 can include ahydrophobic component configured to attract and retain hydrophobic DNAintercalators via hydrophobic interaction from the control sample. Forexample, the hydrophobic component includes octadecylsilyl-basedcomponents (e.g., ODS cartridge) and/or hydrocarbon components (e.g.,C18) or suitable equivalent that can trap and retain DNA intercalatorsfrom the filtered sample. The DNA intercalator can be trapped in thecontrol sample conditioner unit 14 via hydrophobic interaction.

The hydrophobic component that traps the DNA intercalators can bereplaceable or replenishable when the DNA intercalator capacity isreduced. Also, the control sample conditioner unit 14 can include areverse-phase column and associated equipment as the hydrophobiccomponent.

The analytic unit 16 can be configured to receive the test sample andcleansed control sample from the filtration unit 12 (or sample divider22 in FIG. 2) and control sample conditioner unit 14, respectively.Also, the analytic unit 16 can analyze the test sample and cleansedcontrol sample for the presence of one or more DNA intercalators byrunning one or more assays. Additionally, the analytic unit 16 cancalculate the amount of DNA intercalator in the sample.

The analytic unit can be configured to receive the test sample andconditioned control sample, and to contact the test sample andconditioned control sample with double stranded nucleic acids having astrand linked to an ECA substrate. The analytic unit can then detect anelectronic current or change in current that results from the nucleicacids interacting with a DNA intercalator or lack of a DNA intercalatorin the test sample and conditioned control sample; and to obtainelectronic current from the electronic current passed through the testsample and the cleansed control sample.

The analytic unit 16 can include various types of analytical componentsor equipment that operate as described herein, and can include one ormore different types of analytical components. The analytic unit 16 caninclude one or more analytical components selected from NMRspectroscopy, UV-vis, FTIR, mass spectrometry. spectrophotometry,colorimetry, chromatography, electrophoresis, crystallography,microscopy, electrochemistry, titration, gravimetry, qualitativeanalysis, thermal analysis, separation, hybrid techniques (e.g., LC-MS,HPLC-MS, HPLC/ESI-MS, LC-DAD, CE-MS, CE-UV, GC-MS, LC-IR), biochip,chemical arrays, microchip assays (e.g., lab-on-a-chip), and the like aswell as combinations of analytical components. The analytical unit canbe one piece of equipment or it can be an association of two or moresuch of such equipment.

In one embodiment, the analytic unit 16 can include a biochip to testfor the presence or amount of a particular or various DNA intercalators.A biochip (see: Preliminary evaluation of electrochemical PNA array fordetection of single base mismatch mutations, Lab on a chip 1: 61-63,2001; and Microfabricated disposable DNA sensor for detection ofhepatitis B virus DNA, Sensors and Actuators B 46: 220-225, 1998, whichare incorporated herein by specific reference in their entirety) is acollection of miniaturized test sites (e.g., microarrays) arranged on asolid substrate that permits many tests to be performed at the same timein order to achieve higher output and speed. An example of a biochip canbe an electronic chemical array (ECA), which includes aspects ofbiochemistry and semiconductor technologies. Examples of ECAs includewithout limitation the CoulArray® and Coulochem series manufactured byESA, and those prepared by Toshiba Corp. The biochips are an assayplatform that use, in addition to microarray technology, transductionand signal processing technologies to output the results of sensingexperiments. Numerous transduction methods can be employed includingelectronic potential or change, surface plasmon resonance, fluorescence,and chemiluminescence as well as many others. The particular sensing andtransduction techniques chosen depend on factors such as price,sensitivity, and reusability. The actual sensing component (or “chip”)is just one piece of a complete analysis system. Transduction is done totranslate the actual sensing event (i.e., DNA-intercalator binding) intoa format understandable by a computer (e.g., voltage, light intensity,mass, etc.), which then enables additional analysis and processing toproduce a final, human-readable output. A “GeneChip” can containthousands of individual DNA sensors for use in sensing DNAintercalators.

The microarray can include a dense, two-dimensional grid of biosensors.Typically, the sensors are deposited on a flat substrate, which mayeither be passive (e.g. silicon or glass) or active. An active substratecan include integrated electronics or micromechanical devices thatperform or assist signal transduction. Surface chemistry is used tocovalently bind the sensor nucleic acids to the substrate medium.

A core principle behind microarrays is hybridization between two DNAstrands, one being linked to the substrate. The property ofcomplementary nucleic acid sequences to specifically pair with eachother by forming hydrogen bonds between complementary nucleotide basepairs allows for a DNA intercalator to position between the hybridizednucleic acids. The microarrays can use relative quantization in whichthe intensity of a feature when contacted with a sample is compared tothe intensity of the same feature when contacted with the conditionedsample.

A traditional solid-phase microarray can include a collection of orderlymicroscopic “spots”, called features, each with a specific nucleic acidattached to a solid surface, such as glass, plastic or silicon biochip(commonly known gene chip, genome chip, DNA chip or gene array).Thousands of them can be placed in known locations on a single DNAmicroarray.

FIG. 3A is a schematic diagram of an embodiment of an analytic unit 300.The analytic unit 300 includes an inlet port 302 a for the test sampleand an inlet port 302 b for the conditioned control sample. Each inletport 302 a, 302 b is fluidly coupled with a fluid passageway 306 a, 306b that includes one or more valves 308 a, 308 b, 308 c, 308 d, one ormore pumps 310 a, 310 b that can cooperate to pass the test sample andconditioned control sample to an electronic chemical array (ECA) chip312 a, 312 b for analysis. The ECA chips 312 a, 312 b, can include anarray 314 a, 314 b, of analytic modules 316 a, 316 b. Each analyticmodule 316 a, 316 b is associated with one or more electronic pathways318 a, 318 b that can pass an electronic current to one or moreelectronic detectors 320 a, 320 b. The electronic detectors 320 a, 320 bcan be configured detect and/or determine a change in current withrespect to electrodes (not shown) of the analytic modules 316 a, 316 b.The electronic detectors 320 a, 320 b can be associated with datacommunication modules 322 a, 322 b that can be configured to transmitelectronic data from the analytic modules 316 a, 316 b to a remote oronboard data processing unit 324 a, 324 b. The data communicationmodules 322 a, 322 b can also receive operation instruction data fromthe remote or onboard data processing unit 324 a, 324 b and pass thedata to an analytic unit controller 326, which can be configured as acomputing system or microprocessor and associated computing components.The analytic unit controller 326 can be operably coupled with the one ormore inlet ports 302 a, 302 b, valves 308 a, 308 b, 308 c, 308 d, pumps310 a, 310 b, and ECA chips 312 a, 312 b so as to provide operationinstruction data thereto.

In one embodiment, the biochip can include an electronic chemical array(ECA) chip. The electronic chemical array can include a polynucleotidecoupled with a substrate and a complementary polynucleotide thereof thathybridizes with the polynucleotide coupled with the substrate. Thepolynucleotides can be of any size ranging from about 8 nucleotides toabout 100 or more nucleotides, about 10 to about 75 nucleotides, about20 to about 50 nucleotides or about 25 to about 30 nucleotides. Thepolynucleotide coupled to the substrate can have any sequence ofnucleotides, and the complementary polynucleotide can have a sequencethat hybridizes to the polynucleotide coupled to the substrate.

The presence of DNA intercalators can then be identified by a differencein the electronic current of the ECA chip between the conditionedcontrol sample (e.g., having DNA intercalators removed) and the samplesuspected of having the DNA intercalators (e.g., filtered or testsample). Also, the analytic unit 16 can be configured to contact thetest sample and conditioned control sample with double stranded nucleicacids having a strand linked to an electronic chemical array (ECA)reaction surface. The analytic unit 16 can be configured to detect ormeasure an electronic current or change in electronic current throughthe test sample and conditioned control sample. The analytic unit 16 caninclude detection electrodes (e.g., detection electrode, referenceelectrode, and counter electrode) that can obtain electronic data (e.g.,voltage, resistance, current, amplitude, or changes thereof) from theelectronic current detected or measured from the test sample and theconditioned control sample.

The electrodes can operate with the systems and methods described hereinby performing a linear sweep voltammetry (LSV) (see, Preliminaryevaluation of electrochemical PNA array for detection of single basemismatch mutations, Lab on a chip 1: 61-63, 2001). LSV is a voltammetricmethod where the current at the working electrode is measured while thepotential between the working electrode and a reference electrode isswept linearly in time. By LSV, oxidation or reduction of species isregistered as a peak or trough in the current signal at the potential atwhich the species begins to be oxidized or reduced. As such, a change incurrent as detected by the working electrode can be used as theelectronic data.

In another embodiment, the analytic unit 16 can optionally include oneor more of: a port fluidly coupled with the sample divider 22 so as tobe configured to receive the test sample; a port fluidly coupled withthe control sample conditioner unit 14 so as to be configured to receivethe conditioned control sample; one or more sample chambers configuredto receive one or more samples; one or more fluid pathways configured todeliver the test sample and conditioned control sample to the one ormore sample chambers; an integrated or removable ECA; a workingelectrode, counter electrode, and/or reference electrode; a nucleic acidstrand coupled with one of the electrodes; a complementary nucleic acidstrand hybridizable with the electrode-coupled nucleic acid; electroniccomponents configured to cause an electronic current to be passedthrough the electrode-coupled nucleic acid; voltammetry electroniccomponents; electronic components configured to obtain the electroniccurrent; a transmitter to transmit electronic current to the dataprocessing unit; or a receiver to receive instructions from the dataprocessing unit.

FIG. 4 schematically shows a process 100 that can be performed with theECA chip of the analytic unit to determine whether or not a DNAintercalator is present in a sample or the amount thereof. The chip caninclude a nucleic acid 102 a (e.g., DNA) covalently linked with asubstrate 104. The substrate 104 can be associated with a workingelectrode 105 (e.g., copper, titanium, silver, gold or other electrodematerials) such that the working electrode 105 can receive an electroniccurrent from the nucleic acid 102 a through the association of thesubstrate 104 and working electrode. In one aspect, the substrate 104can be the working electrode 105. The nucleic acid 102 a can be arrangedwith a reference electrode 107 and a counter electrode 109 so as to forma circuit capable of having an electronic current. The nucleic acid 102a can be linked to the substrate 104 by a covalent bond or a linker 106or the like. A complementary nucleic acid 102 b can be hybridized withthe nucleic acid 102 a to form a duplex 108. The complementary nucleicacid 102 b can be combined with the nucleic acid 102 a before, during,or after combining a sample with the nucleic acid 102 a by beingintroduced to the ECA chip. Before, during, or after forming the duplex108, the sample containing one or more potential DNA intercalators 110can be introduced to the ECA chip having the duplex 108. The DNAintercalator(s) 110 of the sample can then intercalate with the duplex108 as shown by the intercalation 112. Upon the DNA intercalator 110intercalating with the duplex 108, an electronic current 114 generatedthe duplex 108 can be identified or measured. The electronic current 114can be generated by the intercalation event, which arises from theformation of a kind of charge-transfer complex upon intercalation.

The electronic current 114 can be derived from a redox reaction betweenthe DNA intercalator 110 and the duplex 108. The electronic current 114can depend on the amount of DNA intercalator 110 present in the sample.Each type of DNA intercalator 110 can have a unique electric potential.The amount of electronic current 114 or change in current can vary in apositively correlated manner with the DNA intercalator 110.

The amount of nucleic acid 102 a bound to the substrate 104 (e.g.,optionally conductive material operating as a working electrode) at adefined location can vary widely. For example, the amount can be on theorder of about 10⁻¹⁰ to 10⁻¹⁰; however, other amounts can be used. Thenucleic acid 102 a can also be longer than an 8 mer, such as greaterthan a 12 mer, greater than a 20 mer, or more. An example can be about20 to about a 25 mer nucleic acid. The nucleic acid 102 a can be presentin range from 1×10⁻¹⁸ to 1×10⁻¹⁰, 1×10⁻¹⁶ to 1×10⁻¹², or 1×10⁻¹⁴ to1×10⁻¹³ moles.

FIG. 5 illustrates another embodiment of an ECA 500 with the arrangementof the substrate 104 and the electrodes 105 (working), 107 (reference),109 (counter). As shown, the substrate 104 has a counter electrode 109located directly on its surface. The counter electrode 109 has one ortwo insulators 111 a, 111 b located directly on the surface of thecounter electrode 109. A working electrode 105 is located directly onthe surface of an insulator 111 a in a manner such that the workingelectrode 105 is spaced apart and not touching the counter electrode 109and/or the reference electrode 107. A reference electrode 107 is locateddirectly on the surface of an insulator 111 b in a manner such that thereference electrode 107 is spaced apart from the counter electrode 109and/or working electrode 105. The working electrode 105 can have thenucleic acid 102 a bound directly thereto.

FIGS. 6A-6B illustrate an embodiment of an ECA 600 having a substrate104 having a working electrode 105. The substrate 104 is disposable andconfigured to be received into the analytic unit 16, where the analyticunit includes the reference and/or counter electrodes (not shown) inpredetermined orientations for conducting current analysis and currentchange detection as described herein. As shown, the substrate 104 isglass and approximately 30 mm long, 1.5 mm wide, and 1 mm thick. A layerof titanium 116 is sputtered onto the glass substrate 104 and about 500angstroms to about 50 nm thick. A layer of gold for the workingelectrode 105 is sputtered onto the titanium layer 116 and is about 5000angstroms to about 200 nm thick. An insulator 111 is formed onto thegold working electrode 105 so as to form a gold electrode spot 105 a forreceiving the nucleic acid and form a terminal 105 b that can beelectronically coupled with the analytic unit 16 when received therein.

FIGS. 7A-7B an embodiment of an ECA 700 having a substrate 104configured to be received into the analytic unit 16. As shown, thesubstrate 104 includes a working electrode 105 located on an insulator111 and in proximity to a counter electrode 109 and a referenceelectrode 107. The working electrode 105, reference electrode 107, andcounter electrode 109 are each electronically coupled to a connectingpad 113. The connecting pad 113 is configured to be electronicallycoupled with the analytic unit 16 when the ECA 700 is received therein.

The amount of a DNA intercalator 110 that can be detected can rangethrough parts per trillion (ppt), parts per billion (ppb), parts permillion (ppm), nanomolar (nM), micromolar (mM) and the like. This canallow for the degree of detection sensitivity to DNA intercalators.

For example, if a DNA intercalator 110 is present in the analyticalsample, it will enter a gap between the nucleic acid 102 a and itscomplement 102 b, thereby forming a charge-transfer complex which cangenerate or modulate the electric current being detected. That is, theintercalation event generates or modulates the current 114. For example,in one instance there will be an inherent current from the duplexedpolynucleotide coupled to the substrate due to the electrode system, andthe DNA intercalator changes this current to modulate the electronicdata. In another example, there may not be an inherent current arisingfrom the arrangement of the duplexed polynucleotide and electrodearrangement, and the DNA intercalator generates a current byintercalating with the DNA. The value of the current 114, which isderived from the redox of the DNA intercalator 110, can depend on theamount of DNA intercalator 110 present in the sample for the specificelectric potential of the DNA intercalator unique to a target DNAintercalator. Different DNA intercalators can have different electricpotentials. The amount of the current can vary in an amount thatcorrelates to a particular DNA intercalator. In one instance for aferrocene derivative, the electric potential or varied current can beabout 619 mV.

The nucleic acid-linked electrode can be the working electrode. Acounter electrode and optionally a reference electrode can be included.The counter electrode can be platinum or other similar material. Thereference electrode can be silver/silver-chloride in a suitablebuffer-containing electrolyte. The complementary nucleic acid 102 b canbe included at any time before the sample is contacted to the nucleicacid.

In one example, an equimolar amount of DNA (or more) that has acomplementary sequence with the DNA linked to the chip can be added tothe chip so as to be configured to hybridize therewith.

The analytical unit 16 can optionally include a port (not shown) coupled(e.g., fluidly coupled when sample is a liquid) with the sample divider22 so as to be configured to receive the test sample, and/or a portcoupled with the control sample conditioner unit 14 so as to beconfigured to receive the conditioned control sample. The ports can beany type of sample receiving component.

The analytical unit 16 can optionally include one or more samplechambers (not shown) configured to receive one or more samples. Thechambers can be any type of reaction or assay chamber for analyzing thesample and/or the conditioned sample.

The analytic unit 16 can optionally include one or more pathways (notshown) (e.g., fluid pathway, mechanical mover, etc.) configured todeliver the test sample and conditioned control sample to the one ormore sample chambers.

In still another embodiment, the analytic unit 16 can include anintegrated or removable biochip or ECA reaction component. A biochip orECA reaction component can include a working electrode, counterelectrode, and/or reference electrode so that electronic differencesbetween the sample and conditioned sample can be assessed. The biochipor ECA can include a nucleic acid strand operably coupled with one ofthe electrodes, such that electronic data (e.g., electronic current)related to a DNA intercalator 110 interacting with the nucleic acid 102a can be detected and obtained. The biochip or ECA can also include acomplementary nucleic acid strand hybridizable or hybridized with theelectrode-coupled nucleic acid such that a DNA intercalator 110 can beintercalated between the nucleic acid 102 a and the complementarynucleic acid 102 b to generate or change the electronic current 114.

In still another embodiment, the analytical unit 16 can includevoltammetry electronic components that can perform votammetry methods.Voltammetry is a category of electroanalytical methods used inanalytical chemistry and various industrial processes. In voltammetry,information about an analyte (e.g., DNA intercalator) is obtained bymeasuring the current as the potential is varied. Voltammetryexperiments can investigate the half cell reactivity of an analyte. Mostexperiments control the potential (volts) of an electrode in contactwith the analyte while measuring the resulting current (amperes).Conducting such a votammetry experiment involves at least twoelectrodes, i.e., a working electrode and a reference electrode. Theworking electrode, which makes contact with the analyte, applies thedesired potential in a controlled way and facilitates the transfer ofelectrons to and from the analyte. A counter electrode acts as the otherhalf of the cell. This counter electrode can have a known potential withwhich to gauge the potential of the working electrode, furthermore itcan balance the electrons added or removed by the working electrode. Therole of supplying electrons and referencing potential can be dividedbetween two separate electrodes.

A three electrode detection system can include the working electrode,reference electrode, and counter electrode. The reference electrode canbe a half cell with a known reduction potential. Its only role is to actas reference in measuring and controlling the working electrodespotential. The auxiliary electrode passes the current needed to balancethe current observed at the working electrode. To achieve this current,the auxiliary will often swing to extreme potentials at the edges of thesolvent window, where it oxidizes or reduces the solvent or supportingelectrolyte. These electrodes, the working, reference, and auxiliarymake up the modern three electrode system. For example, a cyclicvoltammogram can be generated.

The analytic unit 16 can include a transmitter (not shown) to transmitelectronic data 114 to the data processing unit 18. Also, the analyticunit 16 can include a receiver (not shown) to receive instructions fromthe data processing unit 18. A transceiver can be included in place of aseparate transmitter and receiver.

The data processing unit 18 can be operably coupled with the analyticunit 16 and configured to compare and determine a difference between theelectronic current 114 from the test sample and the conditioned controlsample. The difference in electronic current 114 can provide anindication of whether or not a DNA intercalator 110 is present in thesample. The processing unit 18 can be any type of computing system withhardware and software that can be used to receive data from the analyticunit 16 and process the data in order to determine the presence oramount of the DNA intercalator 110 in the environmental sample. Thepresence or levels of one or more DNA intercalators 110 can beidentified in the environmental sample by comparing the current 114 orother electronic data of the filtered sample with various standards orwith the conditioned sample (e.g. DNA intercalators removed). Thestandards can be certain electric potential or electronic data, which isunique to a target DNA intercalator 110.

In general, LSV data are affected by measurement conditions. As such,the values obtained by the same analyte during an LSV procedure, suchits peak potential and current, are varied depending upon the conditionused. Therefore, the DNA intercalators that are to be detected with thesystem and methods described herein can be pre-tested in order toidentify their electrochemical characteristics including dose responsesunder conditions of the systems and methods used in the analysis.

For example, the electronic data related to the difference between thesample and control can be current data. When the difference is below a“selected value” (e.g., 20 nA) it is considered that the target DNAintercalator 110 is either not present or present in a minimal or lowrisk amount, when the difference is between selected values (e.g.,between 20 nA and 40 nA) it is considered that the target DNAintercalator 110 is present and at an intermediate or medium risk, andwhen the difference is greater than a selected value (e.g., greater than40 nA) it is considered that the target DNA intercalator 110 is presentat a significant amount that is considered a high risk. The “selectedvalue” is only a rough estimate and determined by pre-testing the DNAintercalator to be detected. The “selected value” is arbitrary anddependent on the type of DNA intercalator, and the configuration of thedetection system as well as on the method used for the detectionprocedure. The “selected values” described herein are relevant toHoechst 33258 (a DNA intercalator) in the water sample by LSV using thesystem and methods described herein.

Also, the data processing unit 18 can include hardware and/or softwareconfigured to operate components of the system. With such hardwareand/or software, the data processing unit 18 can be operably coupledwith the filtration unit 12, sample divider 22, control sampleconditioner unit 14 and the analytic unit 26.

FIGS. 8A-8B illustrate another embodiment of a DNA intercalationdetection system 800 that includes a main housing 802 with a port 801configured to receive a removable cartridge 804. The port 801 of themain housing 802 can include a first connector 810 a and a secondconnector 810 b which are configured to be electronically coupled withelectronic connectors 824 a, 814 b of the removable cartridge 804. Theport 801 of the main housing 802 can also include a suction pump 828configured to be fluidly coupled with a fluid outlet 830 of theremovable cartridge 804. The main housing 802 may optionally include anonboard data processing unit (not shown) or can be operably coupled withan external data processing unit 18.

The removable cartridge 804 can include a sample port 806 fluidlycoupled with a filtration unit 812 that is also fluidly coupled with asample divider 822 that is configured to split a filtered sample. Thesample divider 822 is fluidly coupled with a test sample conditionerunit 813 and a control sample conditioner unit 814. The test sampleconditioner unit 813 is fluidly coupled to a first analytic unit 816 a,and the control sample conditioner unit 814 is fluidly coupled to asecond analytic unit 816 b. An oligonucleotide reservoir 820 is alsofluidly coupled with the first analytic unit 816 a and the secondanalytic unit 816 b. The oligonucleotide reservoir 820 is configured toinclude the antisense nucleic acid 102 b that hybridizes with thesubstrate-bound nucleic acid 102 a. The oligonucleotide reservoir 820can also include a medium, such as a liquid buffer, for moving theoligonucleotide 102 b into the analytic units 816 a, 816 b. The analyticunits 816 a, 816 b are each electronically coupled through electronicpathways 823 a, 623 b to electronic connectors 824 a, 824 b that can beelectronically coupled with the first connector 810 a and secondconnector 810 b of the main housing 812. The removable cartridge 804 canalso include a fluid outlet 830 that is fluidly coupled with the firstanalytic unit 816 a and the second analytic unit 816 b, and isconfigured to be fluidly coupled with the suction pump 828 of the mainhousing 802 when the removable cartridge 804 is received into the port802 of the main housing 802.

The filtration unit 812, sample divider 822, control sample conditionerunit 814, and the analytic units 816 a, 816 b can be configured asdescribed herein. The sample port 806 can be configured as any type offluid sample receiving component, such as a reservoir or container. Thetest sample conditioner unit 813 can be configured similarly as thecontrol sample conditioner unit 814 except that it does not include amaterial (ODS) that binds with a hydrophobic DNA intercalator; insteadthe test sample conditioner unit 813 can include an inert silica gel orother inert medium that does not react or interact with a hydrophobicDNA intercalator.

The removable cartridge 804 can be configured to be disposable. Also,the removable cartridge 804 can be configured as a microfluidic devicewhere the fluid pathways are microchannels and the components aremicrocomponents.

The system can include one or more user interfaces (not shown), whichinclude a user input interface and an output interface. Such userinterfaces can include graphical displays, printer, sound speakers,microphone, keyboards, mouse, light pens, touch screens, buttons, knobs,levers, switches, lights, and the like. For example, each components ofthe system can include separate user interfaces, or a central userinterface can be configured to control some or all of the components.

Embodiments of the system or data processing unit 18 may include orutilize a special purpose or general-purpose computer including computerhardware, as discussed in greater detail below. Embodiments within thescope of the system and data processing unit 18 also include physicaland other computer-readable media for carrying or storingcomputer-executable instructions and/or data structures. Suchcomputer-readable media can be any available media that can be accessedby a general purpose or special purpose computer system.Computer-readable media that store computer-executable instructions arephysical storage media including recordable-type storage media.Computer-readable media that carry computer-executable instructions aretransmission media. Thus, by way of example, and not limitation,embodiments of the system can comprise at least two distinctly differentkinds of computer-readable media: physical storage media andtransmission media.

Physical storage media includes RAM, ROM, EEPROM, CD-ROM or otheroptical disk storage, magnetic disk storage or other magnetic storagedevices, or any other medium which can be used to store desired programcode means in the form of computer-executable instructions or datastructures and which can be accessed by a general purpose or specialpurpose computer.

The system or data processing unit 18 can be linked to a network so thatthe data can be transmitted remotely via wire, optical, wireless or thelike. A “network” is defined as one or more data links that enable thetransport of electronic data between computer systems and/or modulesand/or other electronic devices. When information is transferred orprovided over a network or another communications connection (eitherhardwired, wireless, or a combination of hardwired or wireless) to acomputer, the computer properly views the connection as a transmissionmedium. Transmission media can include a network and/or data links whichcan be used to carry or transport desired program code means in the formof computer-executable instructions or data structures and which can beaccessed by a general purpose or special purpose computer. Combinationsof the above should also be included within the scope ofcomputer-readable media.

However, it should be understood, that upon reaching various computersystem components, program code means in the form of computer-executableinstructions or data structures can be transferred automatically fromtransmission media to physical storage media. For example,computer-executable instructions or data structures received over anetwork or data link can be buffered in RAM within a network interfacecard, and then eventually transferred to computer system RAM and/or toless volatile physical storage media at a computer system. Thus, itshould be understood that physical storage media can be included incomputer system components that also (or even primarily) utilizetransmission media.

The system or data processing unit 18 can include a storage mediumhaving computer-executable instructions for performing the analyticalprotocols and/or data processing as described herein.Computer-executable instructions comprise, for example, instructions anddata which cause a general purpose computer, special purpose computer,or special purpose processing device to perform a certain function orgroup of functions. The computer executable instructions may be, forexample, binaries, intermediate format instructions such as assemblylanguage, or even source code. Although the subject matter has beendescribed in language specific to structural features and/ormethodological acts, it is to be understood that the subject matterdefined in the appended claims is not necessarily limited to thedescribed features or acts described above. Rather, the describedfeatures and acts are disclosed as example forms of implementing theclaims.

Those skilled in the art will appreciate that the data acquisition ordata communication may be practiced in network computing environmentswith many types of computer system configurations, including, personalcomputers, desktop computers, laptop computers, message processors,hand-held devices, multi-processor systems, microprocessor-based orprogrammable consumer electronics, network PCs, minicomputers, mainframecomputers, mobile telephones, PDAs, pagers, routers, switches, and thelike. The data processing or communication may also be practiced indistributed system environments where local and remote computer systems,which are linked (either by hardwired data links, wireless data links,or by a combination of hardwired and wireless data links) through anetwork, both perform tasks. In a distributed system environment,program modules may be located in both local and remote memory storagedevices.

One skilled in the art will appreciate that for processes and methodsdisclosed herein the functions performed in the processes and methodsmay be implemented in differing order. Furthermore, the outlined stepsand operations are only provided as examples, and some of the steps andoperations may be optional, combined into fewer steps and operations, orexpanded into additional steps and operations without detracting fromthe essence of the disclosed embodiments.

The present disclosure is not to be limited in terms of the particularembodiments described in this application, which are intended asillustrations of various aspects. Many modifications and variations canbe made without departing from its spirit and scope, as will be apparentto those skilled in the art. Functionally equivalent methods andapparatuses within the scope of the disclosure, in addition to thoseenumerated herein, will be apparent to those skilled in the art from theforegoing descriptions. Such modifications and variations are intendedto fall within the scope of the appended claims. The present disclosureis to be limited only by the terms of the appended claims, along withthe full scope of equivalents to which such claims are entitled. It isto be understood that this disclosure is not limited to particularmethods, reagents, compounds compositions or biological systems, whichcan, of course, vary. It is also to be understood that the terminologyused herein is for the purpose of describing particular embodimentsonly, and is not intended to be limiting.

In an illustrative embodiment, any of the operations, processes, etc.described herein can be implemented as computer-readable instructionsstored on a computer-readable medium. The computer-readable instructionscan be executed by a processor of a mobile unit, a network element,and/or any other computing device.

There is little distinction left between hardware and softwareimplementations of aspects of systems; the use of hardware or softwareis generally (but not always, in that in certain contexts the choicebetween hardware and software can become significant) a design choicerepresenting cost vs. efficiency tradeoffs. There are various vehiclesby which processes and/or systems and/or other technologies describedherein can be effected (e.g., hardware, software, and/or firmware), andthat the preferred vehicle will vary with the context in which theprocesses and/or systems and/or other technologies are deployed. Forexample, if an implementer determines that speed and accuracy areparamount, the implementer may opt for a mainly hardware and/or firmwarevehicle; if flexibility is paramount, the implementer may opt for amainly software implementation; or, yet again alternatively, theimplementer may opt for some combination of hardware, software, and/orfirmware.

The foregoing detailed description has set forth various embodiments ofthe devices and/or processes via the use of block diagrams, flowcharts,and/or examples. Insofar as such block diagrams, flowcharts, and/orexamples contain one or more functions and/or operations, it will beunderstood by those within the art that each function and/or operationwithin such block diagrams, flowcharts, or examples can be implemented,individually and/or collectively, by a wide range of hardware, software,firmware, or virtually any combination thereof. In one embodiment,several portions of the subject matter described herein may beimplemented via Application Specific Integrated Circuits (ASICs), FieldProgrammable Gate Arrays (FPGAs), digital signal processors (DSPs), orother integrated formats. However, those skilled in the art willrecognize that some aspects of the embodiments disclosed herein, inwhole or in part, can be equivalently implemented in integratedcircuits, as one or more computer programs running on one or morecomputers (e.g., as one or more programs running on one or more computersystems), as one or more programs running on one or more processors(e.g., as one or more programs running on one or more microprocessors),as firmware, or as virtually any combination thereof, and that designingthe circuitry and/or writing the code for the software and or firmwarewould be well within the skill of one of skill in the art in light ofthis disclosure. In addition, those skilled in the art will appreciatethat the mechanisms of the subject matter described herein are capableof being distributed as a program product in a variety of forms, andthat an illustrative embodiment of the subject matter described hereinapplies regardless of the particular type of signal bearing medium usedto actually carry out the distribution. Examples of a signal bearingmedium include, but are not limited to, the following: a recordable typemedium such as a floppy disk, a hard disk drive, a CD, a DVD, a digitaltape, a computer memory, etc.; and a transmission type medium such as adigital and/or an analog communication medium (e.g., a fiber opticcable, a waveguide, a wired communications link, a wirelesscommunication link, etc.).

Those skilled in the art will recognize that it is common within the artto describe devices and/or processes in the fashion set forth herein,and thereafter use engineering practices to integrate such describeddevices and/or processes into data processing systems. That is, at leasta portion of the devices and/or processes described herein can beintegrated into a data processing system via a reasonable amount ofexperimentation. Those having skill in the art will recognize that atypical data processing system generally includes one or more of asystem unit housing, a video display device, a memory such as volatileand non-volatile memory, processors such as microprocessors and digitalsignal processors, computational entities such as operating systems,drivers, graphical user interfaces, and applications programs, one ormore interaction devices, such as a touch pad or screen, and/or controlsystems including feedback loops and control motors (e.g., feedback forsensing position and/or velocity; control motors for moving and/oradjusting components and/or quantities). A typical data processingsystem may be implemented utilizing any suitable commercially availablecomponents, such as those typically found in datacomputing/communication and/or network computing/communication systems.

The herein described subject matter sometimes illustrates differentcomponents contained within, or connected with, different othercomponents. It is to be understood that such depicted architectures aremerely exemplary, and that in fact many other architectures can beimplemented which achieve the same functionality. In a conceptual sense,any arrangement of components to achieve the same functionality iseffectively “associated” such that the desired functionality isachieved. Hence, any two components herein combined to achieve aparticular functionality can be seen as “associated with” each othersuch that the desired functionality is achieved, irrespective ofarchitectures or intermedial components. Likewise, any two components soassociated can also be viewed as being “operably connected”, or“operably coupled”, to each other to achieve the desired functionality,and any two components capable of being so associated can also be viewedas being “operably couplable”, to each other to achieve the desiredfunctionality. Specific examples of operably couplable include but arenot limited to physically mateable and/or physically interactingcomponents and/or wirelessly interactable and/or wirelessly interactingcomponents and/or logically interacting and/or logically interactablecomponents.

With respect to the use of substantially any plural and/or singularterms herein, those having skill in the art can translate from theplural to the singular and/or from the singular to the plural as isappropriate to the context and/or application. The varioussingular/plural permutations may be expressly set forth herein for sakeof clarity.

It will be understood by those within the art that, in general, termsused herein, and especially in the appended claims (e.g., bodies of theappended claims) are generally intended as “open” terms (e.g., the term“including” should be interpreted as “including but not limited to,” theterm “having” should be interpreted as “having at least,” the term“includes” should be interpreted as “includes but is not limited to,”etc.). It will be further understood by those within the art that if aspecific number of an introduced claim recitation is intended, such anintent will be explicitly recited in the claim, and in the absence ofsuch recitation no such intent is present. For example, as an aid tounderstanding, the following appended claims may contain usage of theintroductory phrases “at least one” and “one or more” to introduce claimrecitations. However, the use of such phrases should not be construed toimply that the introduction of a claim recitation by the indefinitearticles “a” or “an” limits any particular claim containing suchintroduced claim recitation to embodiments containing only one suchrecitation, even when the same claim includes the introductory phrases“one or more” or “at least one” and indefinite articles such as “a” or“an” (e.g., “a” and/or “an” should be interpreted to mean “at least one”or “one or more”); the same holds true for the use of definite articlesused to introduce claim recitations. In addition, even if a specificnumber of an introduced claim recitation is explicitly recited, thoseskilled in the art will recognize that such recitation should beinterpreted to mean at least the recited number (e.g., the barerecitation of “two recitations,” without other modifiers, means at leasttwo recitations, or two or more recitations). Furthermore, in thoseinstances where a convention analogous to “at least one of A, B, and C,etc.” is used, in general such a construction is intended in the senseone having skill in the art would understand the convention (e.g., “ asystem having at least one of A, B, and C” would include but not belimited to systems that have A alone, B alone, C alone, A and Btogether, A and C together, B and C together, and/or A, B, and Ctogether, etc.). In those instances where a convention analogous to “atleast one of A, B, or C, etc.” is used, in general such a constructionis intended in the sense one having skill in the art would understandthe convention (e.g., “ a system having at least one of A, B, or C”would include but not be limited to systems that have A alone, B alone,C alone, A and B together, A and C together, B and C together, and/or A,B, and C together, etc.). It will be further understood by those withinthe art that virtually any disjunctive word and/or phrase presenting twoor more alternative terms, whether in the description, claims, ordrawings, should be understood to contemplate the possibilities ofincluding one of the terms, either of the terms, or both terms. Forexample, the phrase “A or B” will be understood to include thepossibilities of “A” or “B” or “A and B.”

In addition, where features or aspects of the disclosure are describedin terms of Markush groups, those skilled in the art will recognize thatthe disclosure is also thereby described in terms of any individualmember or subgroup of members of the Markush group.

As will be understood by one skilled in the art, for any and allpurposes, such as in terms of providing a written description, allranges disclosed herein also encompass any and all possible subrangesand combinations of subranges thereof. Any listed range can be easilyrecognized as sufficiently describing and enabling the same range beingbroken down into at least equal halves, thirds, quarters, fifths,tenths, etc. As a non-limiting example, each range discussed herein canbe readily broken down into a lower third, middle third and upper third,etc. As will also be understood by one skilled in the art all languagesuch as “up to,” “at least,” and the like include the number recited andrefer to ranges which can be subsequently broken down into subranges asdiscussed above. Finally, as will be understood by one skilled in theart, a range includes each individual member. Thus, for example, a grouphaving 1-3 cells refers to groups having 1, 2, or 3 cells. Similarly, agroup having 1-5 cells refers to groups having 1, 2, 3, 4, or 5 cells,and so forth.

From the foregoing, it will be appreciated that various embodiments ofthe present disclosure have been described herein for purposes ofillustration, and that various modifications may be made withoutdeparting from the scope and spirit of the present disclosure.Accordingly, the various embodiments disclosed herein are not intendedto be limiting, with the true scope and spirit being indicated by thefollowing claims.

1. A DNA intercalator detection system, the system comprising: a controlsample conditioner having a DNA intercalator trap member adapted toreceive a control sample and to remove a DNA intercalator from thecontrol sample so as to produce a conditioned control sample; ananalytic unit adapted to receive a test sample and to receive theconditioned control sample from the control sample conditioner, whereinthe analytic unit has one or more electronic chemical arrays (ECAs); anda data processing unit adapted to compare and to determine a differencebetween an electronic current of the test sample with an electroniccurrent of the conditioned control sample from the one or more ECAs,said difference providing an indication of whether or not a DNAintercalator is present in the test sample.
 2. The system of claim 1,further comprising a filtration unit, wherein the filtration unitincludes filter components adapted to separate environmental and/orbiological materials from an environmental sample.
 3. The system ofclaim 1, wherein the DNA intercalator detection system is portable. 4.The system of claim 2, wherein the filtration unit includes a filterhaving polyvinylidene difluoride (PVDF).
 5. The system of claim 2,further comprising: a sample divider adapted to receive a filteredsample from the filtration unit, and to split the filtered sample intotwo or more portions.
 6. The system of claim 1, wherein the DNAintercalator trap member includes a hydrophobic component adapted toattract and retain hydrophobic DNA intercalators from the controlsample.
 7. The system of claim 6, wherein the hydrophobic componentincludes octadecylsilyl-based components and/or hydrocarbon components.8. The system of claim 1, wherein the one or more ECAs include one ormore polynucleotides coupled to a substrate.
 9. The system of claim 8,further comprising the analytic unit having one or more electrodesadapted to detect electronic current in response to a DNA intercalatorintercalating with the one or more polynucleotides that are hybridizedwith complementary polynucleotides.
 10. A method for detecting a DNAintercalator in an environmental sample, the method comprising:splitting an environmental sample into a test sample and a controlsample; conditioning the control sample such that one or more DNAintercalators are removed therefrom to form a conditioned controlsample; contacting the test sample with double stranded nucleic acidshaving a strand linked to an electronic chemical array (ECA); contactingthe conditioned control sample with double stranded nucleic acids havinga strand linked to an ECA; detecting an electronic current generated bythe test sample and an electronic current generated by the conditionedcontrol sample; and determining a difference between the electroniccurrent of the test sample and the conditioned control sample, saiddifference providing an indication of whether or not a DNA intercalatoris present in the test sample.
 11. The method of claim 10, furthercomprising collecting the environmental sample from a location andperforming the method at the location.
 12. The method of claim 10,further comprising filtering the environmental sample, the test sample,and/or the control sample, wherein filtering comprises removingbiological materials.
 13. The method of claim 10, further comprisingcontacting one or more portions of the control sample to a DNAintercalator trap member adapted to attract and retain DNA intercalatorstherefrom.
 14. The method of claim 10, comprising measuring theelectronic currents of the test sample and conditioned control sample.15. The method of claim 10, comprising hybridizing a nucleic acid strandcoupled to the ECA with its complementary nucleic acid.
 16. The methodof claim 10, comprising conducting a voltammetry protocol.
 17. Themethod of claim 10, further comprising processing the electronic data inorder to determine a difference between the electronic data from thetest sample and the conditioned control sample.
 18. The method of claim10, further comprising determining an amount or health risk of a DNAintercalator.
 19. The method of claim 18, wherein the determining thehealth risk is performed by identifying the difference between theelectronic current of the test sample with the electronic current of theconditioned control sample.
 20. A method for detecting a DNAintercalator in an environmental sample, the method comprising:introducing the environmental sample into a DNA intercalator detectionsystem, said DNA intercalator detection system including: a filtrationunit; a control sample conditioner; an analytic unit; and a dataprocessing unit; filtering the environmental sample with the filtrationunit so as to remove environmental substances from the environmentalsample and provide a filtered sample; conditioning a portion of thefiltered sample with the control sample conditioner so as to remove oneor more DNA intercalators and form a conditioned control sample;contacting, separately in the analytic unit, a test sample and theconditioned control sample with double stranded nucleic acids eachhaving a strand linked to an electronic chemical array (ECA); obtainingelectronic data from an electronic current of the test sample and theconditioned control sample when in contact with the double strandednucleic acids; and determining a difference between the electronic datafrom the test sample and the conditioned control sample with the dataprocessing unit, said difference providing an indication of whether ornot a DNA intercalator is present in the sample.